4.7 Article

An effective biphase system accelerates hesperidinase-catalyzed conversion of rutin to isoquercitrin

Journal

SCIENTIFIC REPORTS
Volume 5, Issue -, Pages -

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/srep08682

Keywords

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Funding

  1. China Postdoctoral Science Foundation [2013T60505, 2012M510124]
  2. Postdoctoral Science Foundation Funded Project of Jiangsu University [1143002085]
  3. Qing Lan Project of Jiangsu Province
  4. Start Project for Introduced Talent at Jiangsu University of Science and Technology [35211002]

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Isoquercitrin is a rare, natural ingredient with several biological activities that is a key precursor for the synthesis of enzymatically modified isoquercitrin (EMIQ). The enzymatic production of isoquercitrin from rutin catalyzed by hesperidinase is feasible; however, the bioprocess is hindered by low substrate concentration and a long reaction time. Thus, a novel biphase system consisting of [Bmim][BF4]: glycine-sodium hydroxide (pH 9) (10:90, v/v) and glyceryl triacetate (1:1, v/v) was initially established for isoquercitrin production. The biotransformation product was identified using liquid chromatography-mass spectrometry, and the bonding mechanism of the enzyme and substrate was inferred using circular dichroism spectra and kinetic parameters. The highest rutin conversion of 99.5% and isoquercitrin yield of 93.9% were obtained after 3 h. The reaction route is environmentally benign and mild, and the biphase system could be reused. The substrate concentration was increased 2.6-fold, the reaction time was reduced to three tenths the original time. The three-dimensional structure of hesperidinase was changed in the biphase system, which a-helix and random content were reduced and beta-sheet content was increased. Thus, the developed biphase system can effectively strengthen the hesperidinase-catalyzed synthesis of isoquercitrin with high yield.

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